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棒状体激酶家族蛋白17诱导的LMH细胞转录变化

Transcriptional changes in LMH cells induced by rhoptry kinase family protein 17.

作者信息

Meng Yi-Jing, Mu Bing-Jin, Liu Xiao-Xin, Yu Lin-Mei, Zheng Wen-Bin, Xie Shi-Chen, Gao Wen-Wei, Zhu Xing-Quan, Liu Qing

机构信息

College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong, China.

Key Laboratory of Veterinary Public Health of Higher Education of Yunnan Province, College of Veterinary Medicine, Yunnan Agricultural University, Kunming, China.

出版信息

Front Vet Sci. 2022 Aug 9;9:956040. doi: 10.3389/fvets.2022.956040. eCollection 2022.

Abstract

Though a number of rhoptry kinase family proteins have been identified, little is known about their molecular functions. In the present study, the gene fragment encoding the matured peptide of rhoptry kinase family protein 17 (EtROP17) was used to construct a recombinant vector, followed by transfection into leghorn male hepatoma (LMH) cells. Then, the transcriptional changes in the transfected cells were determined by RNA-seq. The expression of EtROP17 in LMH cells was validated by both Western blot and indirect immunofluorescence analysis. Our analysis showed that EtROP17 altered the expression of 309 genes (114 downregulated genes and 195 upregulated genes) in LMH cells. The quantitative real-time polymerase chain reaction (qRT-PCR) results of the selected differentially expressed genes (DEGs) were consistent with the RNA-seq data. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that DEGs were significantly enriched in nine pathways, such as toll-like receptor signaling pathway, ECM-receptor interaction, intestinal immune network for IgA production and focal adhesion. These findings reveal several potential roles of EtROP17, which contribute to understanding the molecular mechanisms underlying the host-parasite interplay.

摘要

尽管已经鉴定出许多棒状体激酶家族蛋白,但其分子功能却知之甚少。在本研究中,编码棒状体激酶家族蛋白17(EtROP17)成熟肽的基因片段被用于构建重组载体,随后转染至来航鸡雄性肝癌(LMH)细胞中。然后,通过RNA测序确定转染细胞中的转录变化。通过蛋白质免疫印迹和间接免疫荧光分析验证了EtROP17在LMH细胞中的表达。我们的分析表明,EtROP17改变了LMH细胞中309个基因的表达(114个基因下调,195个基因上调)。所选差异表达基因(DEG)的定量实时聚合酶链反应(qRT-PCR)结果与RNA测序数据一致。京都基因与基因组百科全书(KEGG)分析表明,差异表达基因在九个途径中显著富集,如Toll样受体信号通路、细胞外基质-受体相互作用、IgA产生的肠道免疫网络和粘着斑。这些发现揭示了EtROP17的几个潜在作用,有助于理解宿主-寄生虫相互作用的分子机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fac3/9395702/dda603a74214/fvets-09-956040-g0001.jpg

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